ProLong® antifade reagents  

ProLong® antifade mountants suppress photobleaching and preserve the signals of your fluorescently labeled target molecules for long-term storage and analysis. ProLong® reagents cure within 24 hours, forming a semi-rigid gel with a refractive index of 1.46. Mounted samples can be saved for months with appropriate storage. Key benefits of ProLong® antifade reagents include:

  • Inhibit photobleaching across the spectrum, even after prolonged storage
  • Hard-setting mountants
  • Available with or without DAPI
  • Ready-to-use bench-top formulations
  • Refractive index 1.42 after overnight curing, increasing to 1.46 over several days

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Choose the antifade reagent that matches your experiment

Performance of ProLong® antifade reagents

ProLong® Gold antifade reagent performance

ProLong® Gold antifade reagent has become a widely used antifade reagent for Molecular Probes® high-performance fluorescent dyes and stains because it preserves signal across the entire visible spectrum and causes little or no quenching of the initial signal. ProLong® Gold is room-temperature stable—keep it on the bench, ready to use in your next imaging experiment.

ProLong® Diamond antifade reagent performance

ProLong® Diamond antifade reagent delivers all the performance of ProLong® Gold reagent with Alexa Fluor® dyes and provides additional protection for traditional dyes and fluorescent proteins. ProLong® Diamond reagent is a superior antifade and mountant, offering protection across the visible spectrum. Keep ProLong® Diamond reagent refrigerated for the best performance.

Performance of ProLong® antifade reagents

A 60-second time-lapse showing the enhanced resistance to photobleaching afforded by ProLong® antifade reagents. Fixed HeLa cells were labeled with fluorescein phalloidin and mounted in ProLong® Diamond reagent, ProLong® Gold reagent, or 50% PBS/glycerol. Images were acquired at 12-second intervals using a 20x objective with continuous illumination from a standard 100-watt Hg-arc lamp.

ProLong® antifade reagents

Photobleaching resistance for fluorescent dyes and proteins with ProLong® antifade reagents. Photobleaching resistance was quantified on a LSM 710 (Zeiss) confocal microscope. HeLa or U2OS cells were stained and mounted using standard ICC protocols. Five regions within three fields of view were scanned 50x with a 1.58 μs dwell time per pixel. Excitation wavelength and intensity were optimized for the fluorophore being measured.

ProLong® reagent gallery

ProLong® Diamond antifadeEnlarge image
ProLong® Diamond antifadeEnlarge image
ProLong® Diamond antifadeEnlarge image
ProLong® Diamond antifadeEnlarge image
ProLong® Diamond antifadeEnlarge image
ProLong® Diamond antifadeEnlarge image

Find the right ProLong® antifade reagent

 
ProLong® Gold antifade reagent
ProLong® Diamond antifade reagent
Alexa Fluor® dyes
Yes
Yes
Yes
Yes
Traditional dyes & FPs
No
No
Yes
Yes
RT storage
Yes
Yes
No
No
Counterstain
No
DAPI
No
DAPI
Format 5 x 2 mL 10 mL 5 x 2 mL 10 mL
5 x 2 mL
5 x 2 mL
Cat. No. P36934 P36930 P36935 P36931

Performance across the spectrum

ProLong® Diamond reagent is a superior antifade and mountant, offering protection across the visible spectrum and exhibiting less initial quenching of the fluorescent signal. It provides additional protection for traditional dyes such as FITC and fluorescent proteins such as GFP. In a comparison of commercial hard-mount antifade reagents in a standardized experiment, ProLong® mountants outperformed other products across the analysis spectrum.

Comparison of degree of photobleaching protection by ProLong® antifades for various fluorescent dyes

Fluorescent dye
Ex/Em (nm)
Resistance to photobleaching*

ProLong® Gold

ProLong® Diamond
VECTASHIELD® HardSet
Mounting Medium
Hoechst®
350/461
100%
100%
90%
DAPI
345/455
90%
90%
90%
BODIPY® FL
505/513
70%
100%
50%
Alexa Fluor® 488
495/519
80%
100%
90%
GFP
488/510
40%
90%
90%
Fluorescein
494/518
80%
90%
90%
Cy®3
550/570
80%
80%
80%
Alexa Fluor® 546
556/575
100%
100%
100%
Tetramethylrhodamine
555/580
80%
80%
80%
Alexa Fluor® 555
555/565
90%
100%
90%
Alexa Fluor® 568
578/603
70%
90%
90%
Texas Red®
595/615
90%
90%
90%
Alexa Fluor® 594
590/617
100%
100%
90%
TO-PRO®-3
642/661
90%
90%
90%
Alexa Fluor® 647
652/668
100%
90%
0%
Cy®5
650/670
100%
100%
90%
*The degree of photobleaching protection is compiled from a combination of bright field exposure and confocal scanning data to represent typical user experience in fluorescence imaging applications.
VECTASHIELD is a registered trademark of Vector Laboratories, Inc., Burlingame, CA. Hoechst is a trademark of Hoechst, GmbH. Cy is a trademark of GE Healthcare UK, Ltd.

Green chemistry

green chemistry

 

For added convenience and a reduction in the environmental impact associated with cold storage requirements, our antifade reagents are provided in dropper bottles with storage right at your bench. In addition, these reagents are formulated with reduced sodium azide to help minimize environmental and health hazard exposure risks. Learn more about these green chemistry initiatives

For Research Use Only. Not for use in diagnostic procedures.